茶树休眠相关基因CsAGL8的克隆与表达分析  

Cloning and Expression Analysis of a MADS-box Gene CsAGL8 in the Tea Plant(Camellia sinensis)

作　　者：胡小莉 付一丹 崔懂 夏心杰 陈盛相[1,2] 唐茜 谭礼强[1,2] HU Xiaoli;FU Yidan;CUI Dong;XIA Xinjie;CHEN Shengxiang;TANG Qian;TAN Liqiang(College of Horticulture,Sichuan Agricultural University,Chengdu 611130,China;Tea Refining and Innovation Key Laboratory of Sichuan Province,Chengdu 611130,China)

机构地区：[1]四川农业大学园艺学院,四川成都611130 [2]精制川茶四川省重点实验室,四川成都611130

出　　处：《云南农业大学学报（自然科学版）》2023年第6期1032-1040,共9页Journal of Yunnan Agricultural University:Natural Science

基　　金：国家自然科学基金项目(31800589);大学生创新性实验计划项目(202010626132);四川省科技计划项目(2022JDRC0035)。

摘　　要：【目的】克隆茶树CsAGL8基因序列,并探究该基因与茶树休眠的关系。【方法】以峨眉问春等茶树品种的休眠芽为材料,采用RT-PCR扩增MADS-box转录因子家族基因CsAGL8,并进行BLAST比对等生物信息学分析;采用实时荧光定量PCR检测CsAGL8在茶树8个组织和越冬期间的相对表达量;遗传转化拟南芥,对转基因植株进行表型观测及开花相关基因分析。【结果】CsAGL8的CDS全长为720 bp,编码239个氨基酸,含有高度保守的MADS-box结构域和半保守的K-box结构域,与拟南芥的AtAGL8相似性最高。CsAGL8编码区表现出一定的多态性,启动子区域存在较多的光、激素和胁迫响应等顺式作用元件。表达分析表明:CsAGL8在茶树不同组织中均有表达,在休眠顶芽中的表达量最高;越冬期间,CsAGL8的表达量呈先升高后降低的趋势,在特早生品种峨眉问春芽中的相对表达量显著低于对照品种,且下调时间早于对照品种。过表达CsAGL8,拟南芥转基因植株会早花10~15 d。【结论】CsAGL8可能参与了茶树越冬休眠调控。[Purpose]To clone the CsAGL8 gene sequence of tea plant and to explore the relation-ship between this gene and tea plant dormancy.[Methods]The MADS-box transcription factor family gene CsAGL8 was cloned from the dormant buds of the tea cultivar‘Emei Wenchun’by RT-PCR,and bioinformatics analysis such as BLAST comparison was performed.The relative expres-sion levels in eight tissue samples and during overwintering were determined by qPCR.Genetic trans-formation of Arabidopsis thaliana,to observe the phenotype of transgenic plants and analyze the flowering-related genes.[Results]The CDS of CsAGL8 was 720 bp,which encoded 239 amino acids,and contained a highly conserved MADS-box domain and a semi-conserved K-box domain.CsAGL8 showed the highest similarity to AtAGL8 from A.thaliana.The coding region of CsAGL8 showed polymorphisms,and many cis-acting elements such as light,hormone,and stress response were found in the promoter region.Expression analysis showed that CsAGL8 was expressed in differ-ent tissues of tea plants,and the highest expression was detected in the dormant terminal buds.Dur-ing the overwintering process,the expression of CsAGL8 increased first and then decreased.Com-pared with the regular tea cultivar,the expression levels of CsAGL8 were not only significantly lower,but also down-regulated earlier in the buds of the extra-early cultivar‘Emei Wenchun’.Overexpres-sion of CsAGL8 in A.thaliana promoted early flowering for 10-15 days.[Conclusion]CsAGL8 may be involved in the regulation of winter dormancy in tea plants.

关 键 词：茶树 MADS-BOX 基因克隆 表达分析 休眠 

分 类 号：S571.101[农业科学—茶叶生产加工]